Ab217876

The cells were collected and lysed in the lysis buffer which was mixed with 0.1% sodium dodecyl sulfate (SDS), 50 mM TrisHCl, sodium deoxycholate, 150 mM NaCl, 1% NP-40, and a protease inhibitor (PH = 7.5). Then, the protein samples were loaded to 8% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride membrane (PVDF). After blocking with 5% non-fat dry milk for 1 h, the membranes were incubated with the specific primary antibody overnight. After washing in TBST for three times in 15 min, the membranes were then incubated with the secondary antibodies (peroxidase-labeled anti-mouse and anti-rabbit antibodies) for 1 h. Briefly, the primary antibodies were utilized at a dilution of 1:2,000, while secondary antibodies were diluted with 1:5,000. After washing in TBST for three times in 15 min, the membranes were finally visualized in an ECL + plus^TM Western Blotting system kit (cat. no. RPN2232; Amersham). The detailed information of the primary antibodies was summarized as follows: KAT2A (abcam, ab217876); MCT1 (abcam, ab90582); β-Actin (CST#4970).

HepG2 and L02 cells were cultured in 100-mm culture dishes and crosslinked with 1% formaldehyde. Then cells were lysed with cell lysis buffer (50 mM Tris-HCl, 5 mM EDTA, 150 mM NaCl, 1% Triton X-100, 0.1% Deoxycholate and PIC). Next, cells were sonicated and the supernatants were treated with EP300 (Abcam, ab14984), GCN5 (Abcam, ab217876) and IgG (Abcam, ab190475; ab172730) primary antibody for 2 h followed by rotation with 25ul dynabeads protein G for 2 more hours. The DNA fractions were purified and used for next qPCR. Primers are as follows: GPNCA-pro-1 forward and reverse primers, 5′-CGCCTTGCACTTCCCCACT-3′ and 5′-TCCCAGACGCCTGTTACGC-3′; GPNCA-pro-2 forward and reverse primers, 5′-TTTGTTTGCGGCTCCTCCC-3′ and 5′-TTGCCATTGTCGAATGTCTCC-3′.

Cells were lysed in RIPA (CWBio, Beijing, China) containing protease inhibitors, on ice. The lysates were mixed with loading buffer and denatured at 100 °C for 10 min. The products were then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, Massachusetts, USA). The membranes were incubated with primary antibodies [TRIM22 (Abcam, ab224059), KAT2A (CST, ab217876), FLAG (Abcam, ab205606), Myc (Abcam, ab32072), Ub (Abcam, ab140601) and GADPH (Affinity, 1:1000)] and then the secondary antibody, which was conjugated to horseradish peroxidase (HRP;anti-rabbit IgG/anti-mouse IgG, CST, 1:5000). Immunoreactive bands were visualized using Image Lab after the addition of luminol-based chemiluminescent substrate (ECL; Millipore). The immunoblotting results were analyzed using Image Lab software.